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Mindin is important in broad spectrum of immune responses. On the other hand, we previously reported that mindin attenuated human colon cancer development by blocking angiogenesis through Egr-1-mediated regulation. However, the mice original mindin directly suppressed the syngenic colorectal cancer (CRC) growth in our recent study and we aimed to further define the role of mindin during CRC development in mice. We established the mouse syngeneic CRC CMT93 and CT26 WT cell lines with stable mindin knock-down or overexpression. These cells were also subcutaneously injected into C57BL/6 and BALB/c mice as well as established a colitis-associated colorectal cancer (CAC) mouse model treated with lentiviral-based overexpression and knocked-down of mindin. Furthermore, we generated mindin knockout mice using a CRISPR-Cas9 system with CAC model. Our data showed that overexpression of mindin suppressed cell proliferation in both of CMT93 and CT26 WT colon cancer cell lines, while the silencing of mindin promoted in vitro cell proliferation via the ERK and c-Fos pathways and cell cycle control. Moreover, the overexpression of mindin significantly suppressed in vivo tumour growth in both the subcutaneous transplantation and the AOM/DSS-induced CAC models. Consistently, the silencing of mindin reversed these in vivo observations. Expectedly, the tumour growth was promoted in the CAC model on mindin-deficient mice. Thus, mindin plays a direct tumour suppressive function during colon cancer progression and suggesting that mindin might be exploited as a therapeutic target for CRC.
Assuntos
Neoplasias do Colo/genética , Proteínas da Matriz Extracelular/genética , Genes Supressores de Tumor/fisiologia , Sistema de Sinalização das MAP Quinases/genética , Transdução de Sinais/genética , Animais , Ciclo Celular/genética , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/genética , Colite/genética , Colite/patologia , Colo/patologia , Neoplasias do Colo/patologia , Modelos Animais de Doenças , Progressão da Doença , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Células RAW 264.7RESUMO
OBJECTIVES: This study aimed to validate the 8th edition of American Joint Committee on Cancer/Union for International Cancer Control (AJCC/UICC) TNM staging system for nasopharyngeal carcinoma (NPC) in non-endemic region. MATERIALS AND METHODS: We recruited 607 patients with histology-proven, previously untreated, non-metastatic NPC treated by intensity-modulated radiotherapy (IMRT) at our center. Harrell's concordance index (c-index) and Akaike information criterion (AIC) were applied to compare the prognostic discrimination between the 7th and 8th edition staging system. RESULTS: For T category, the local recurrence-free survival (LRFS) Kaplan-Meier curves of T1, T2 and T3 were well separated in the 8th edition; however, LRFS did not significantly differ between T3 and T4 (Pâ¯=â¯0.166). Moreover, the 7th edition achieved higher c-index (0.702 [95% CI, 0.618-0.787] vs. 0.685 [95% CI, 0.604-0.767]) and lower AIC (766.1 vs. 770.8) than 8th edition for LRFS. With regard to N category, the 8th edition achieved higher c-index (0.796 [95% CI, 0.749-0.843] vs. 0.751 [95% CI, 0.696-0.805]) and lower AIC (1439.4 vs. 1471.9) for distant metastasis-free survival. In terms of overall stage, the 8th edition also had higher c-index (0.798 [95% CI, 0.753-0.844] vs. 0.721 [95% CI, 0.672-0.770]) and lower AIC (1963.9 vs. 2007.2) compared with the 7th edition for overall survival. Furthermore, interval validation by bootstrapping the sample randomly for ~100-1000 times also validated above findings. CONCLUSION: The 8th edition of AJCC/UICC TNM staging system achieved significantly better prognostic discrimination than the 7th edition with regard to N category and overall stage but not T category.
Assuntos
Carcinoma Nasofaríngeo/diagnóstico , Estadiamento de Neoplasias/métodos , Guias de Prática Clínica como Assunto , Terapia Combinada , Gerenciamento Clínico , Feminino , Seguimentos , Humanos , Masculino , Imagem Multimodal/métodos , Carcinoma Nasofaríngeo/epidemiologia , Carcinoma Nasofaríngeo/mortalidade , Carcinoma Nasofaríngeo/terapia , Estadiamento de Neoplasias/normas , Prognóstico , Reprodutibilidade dos Testes , Resultado do TratamentoRESUMO
Mindin has a broad spectrum of roles in the innate immune system, including in macrophage migration, antigen phagocytosis and cytokine production. Mindin functions as a pattern-recognition molecule for microbial pathogens. However, the underlying mechanisms of mindin-mediated phagocytosis and its exact membrane receptors are not well established. Herein, we generated mindin-deficient mice using the CRISPR-Cas9 system and show that peritoneal macrophages from mindin-deficient mice were severely defective in their ability to phagocytize E coli. Phagocytosis was enhanced when E coli or fluorescent particles were pre-incubated with mindin, indicating that mindin binds directly to bacteria or non-pathogen particles and promotes phagocytosis. We defined that 131 I-labelled mindin binds with integrin Mac-1 (CD11b/CD18), the F-spondin (FS)-fragment of mindin binds with the αM -I domain of Mac-1 and that mindin serves as a novel ligand of Mac-1. Blockade of the αM -I domain of Mac-1 using either a neutralizing antibody or si-Mac-1 efficiently blocked mindin-induced phagocytosis. Furthermore, mindin activated the Syk and MAPK signalling pathways and promoted NF-κB entry into the nucleus. Our data indicate that mindin binds with the integrin Mac-1 to promote macrophage phagocytosis through Syk activation and NF-κB p65 translocation, suggesting that the mindin/Mac-1 axis plays a critical role during innate immune responses.
Assuntos
Proteínas da Matriz Extracelular/metabolismo , Antígeno de Macrófago 1/metabolismo , Macrófagos/citologia , Macrófagos/metabolismo , Fagocitose , Receptores de Reconhecimento de Padrão/metabolismo , Quinase Syk/metabolismo , Fator de Transcrição RelA/metabolismo , Animais , Sequência de Bases , Núcleo Celular/metabolismo , Células HEK293 , Humanos , Antígeno de Macrófago 1/química , Camundongos , Camundongos Knockout , Fosforilação , Ligação Proteica , Domínios Proteicos , Transporte Proteico , Células RAW 264.7RESUMO
OBJECTIVE: To investigate the role of SRC kinase inhibitor PP2 in drug resistance to adriamycin (ADM) in breast cancer cells and invasion, metastasis of cells. METHODS: MTT assay was used to detect the inhibitory effect of ADM on MCF-7 and MCF-7/ADM cells. The 50% inhibitory concentration (IC50) and resistance index (RI) of cells were calculated. The expression of MDR1, connexin 43 (Cx43) and SRC proteins in breast cancer cells were detected by Western blot assay. Transwell experiment and cell scratch test were used to determine the invasion and migration of cells respectively [MCF-7, MCF-7/ADM, PP2 (1, 2, 4 µmol/L)]. Standard colony formation assay was used to detect the cytotoxicity effect of 4 µmol/L PP2 pretreatment on ADM. RESULTS: ADM inhibited the proliferation of MCF-7 more than MCF-7/ADM cells (P<0.01). The IC50 of MCF-7/ADM cells was 24.55 µmol/L, the IC50 of MCF-7/ADM cells was 770.57 µmol/L, the RI was 31. Compared with MCF-7 cells, expressions of the multidrug resistance proteins MDR1 and SRC were significantly increased (P<0.01). The invasion and migration ability of the MCF-7/ADM cells was stronger than that of the sensitive cells (P<0.01). When MCF-7/ADM was exposed to SRC inhibitor PP2, the invasion and metastasis ability of cells were inhibited (P<0.01) and the rate of colony formation was decreased, that is, more sensitivity to ADM (P<0.01). CONCLUSION: The resistance of MCF-7 to ADM is accompanied by increased expression of SRC. SRC inhibitor PP2 can reduce the cell resistance, ability of invasion and metastasis.
Assuntos
Neoplasias da Mama/enzimologia , Doxorrubicina/metabolismo , Resistencia a Medicamentos Antineoplásicos , Pirimidinas/farmacologia , Quinases da Família src/metabolismo , Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Conexina 43/metabolismo , Humanos , Células MCF-7 , Invasividade Neoplásica , Metástase NeoplásicaRESUMO
Background: This study aimed to compare concurrent chemoradiotherapy (CCRT) plus cetuximab (C) with CCRT alone in locoregionally advanced nasopharyngeal carcinoma(NPC). Methods: A total of 682 locoregionally advanced NPC patients who had undergone chemoradiotherapy with or without cetuximab were included. Propensity score-matching method was used to match patients. Progression-free survival (PFS), overall survival (OS), locoregional relapse-free survival (LRFS), and distant metastasis-free survival (DMFS) were compared between the two treatment arms. Results: After matching, 225 patients were identified for the analysis. Compared to CCRT, CCRT plus C was associated with significantly improved 3-year PFS (83.7% vs 71.9%, P = 0.036), LRFS (98.6% vs 90.2%, P = 0.034) but not OS (91.4% vs 85.4%, P = 0.117). Among patients with T4 and/or N3 category, CCRT plus C significantly prolonged 3-year PFS (81.0% vs 61.4%, P = 0.022) and increased 3-year OS (88.0% vs 77.9%, P = 0.086). No significant differences were observed between CCRT plus C and CCRT alone groups with regard to 3-year PFS, OS, LRFS and DMFS rates in stage III patients. Acute oral and oropharyngeal mucositis during radiotherapy were more common in the CCRT plus C than that in CCRT, but late toxicities were comparable. Conclusions: This study reveals that patients with locoregionally advanced NPC could benefit from the addition of cetuximab to CCRT, and this therapeutic gain mainly originated from T4 and/or N3 subgroup although suffering more acute moderate to severe toxicities.
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OBJECTIVE: C1-C2 pedicle screw fixation has become popular for providing excellent bony purchase and avoiding neurovascular complications. However, it may be technically challenging in children. The objective of this study is to investigate the safety and efficacy of C1-C2 pedicle screw fixation for atlantoaxial dislocation (AAD) in pediatric patients younger than 5 years and to evaluate the preliminary clinical and radiographic results. METHODS: During a 7-year period, 15 patients with a mean age of 3.4 years (range, 2-5 years) underwent C1-C2 pedicle screw fixation for AAD; at least 1 C1 pedicle screw was incorporated as part of the posterior atlantoaxial fusion construct. The cause, surgical technique, instrumentation, and clinical and radiographic results were analyzed. RESULTS: Five patients had preoperative neurologic deficits and no neurovascular injury occurred during surgery. Anterior release using a retropharyngeal approach was performed in 4 cases. Fixation of 55 C1 and C2 pedicle screws was performed successfully without neurovascular complications. Anatomic and partial reductions occurred in 12 and 3 cases, respectively. Solid fusion was achieved in 14 patients (96.9%) during a mean follow-up of 37.6 months (range, 12-111 months). Two patients (13%) experienced complications: one had prolonged immobilization for a loose C1 pedicle screw, and one had unintended fusion caused by allograft absorption. All patients showed radiographic stability and symptom resolution. CONCLUSIONS: C1-C2 pedicle screw fixation for AAD is safe and effective even in children younger than 5 years.
Assuntos
Articulação Atlantoaxial/lesões , Articulação Atlantoaxial/cirurgia , Luxações Articulares/cirurgia , Parafusos Pediculares , Fusão Vertebral , Articulação Atlantoaxial/diagnóstico por imagem , Vértebra Cervical Áxis/diagnóstico por imagem , Vértebra Cervical Áxis/cirurgia , Atlas Cervical/diagnóstico por imagem , Atlas Cervical/cirurgia , Pré-Escolar , Estudos de Viabilidade , Feminino , Seguimentos , Humanos , Luxações Articulares/complicações , Luxações Articulares/diagnóstico por imagem , Masculino , Complicações Pós-Operatórias/diagnóstico por imagem , Estudos Retrospectivos , Resultado do TratamentoRESUMO
OBJECTIVES: Intensity-modulated radiotherapy (IMRT) has been applied in nasopharyngeal carcinoma (NPC) for nearly twenty years, while little is known about the ten-year survival outcomes. This study aimed at evaluating the 10-year survival outcomes for patients with NPC receiving IMRT. MATERIALS AND METHODS: Data on 614 patients with newly diagnosed, non-disseminated NPC treated by IMRT between 2004 and 2008 were retrospectively reviewed. Survival outcomes stratified by tumor stage were compared. RESULTS: The median follow-up duration was 112.7months (range, 7.6-156.8months) for the entire cohort. The 10-year local relapse-free survival rates for T1, T2 and T3 were 94.2%, 92.5% and 91.4% (P>0.05), respectively, and significantly higher than that of T4 disease (79.3%, P<0.05 for all rates). As N category increased from N0 to N3, the 10-year distant metastasis-free survival rates significantly decreased accordingly (P<0.01 for all rates). Furthermore, the 10-year overall survival rates were 100%, 87.1%, 75.5% and 55.6% for stage I, II, III and IV, respectively (P<0.05 except stage I and II). Multivariate analysis established tumor stage and age as independent prognostic factors. Late toxicities were assessable for 495 (80.6%) patients and most were Grade I/II damages. Xerostomia (387 of 489, 79.1%) and hearing impairment (212 of 495, 42.8%) remained the most troublesome. CONCLUSION: IMRT could achieve satisfactory survival outcomes for NPC patients with acceptable late toxicities. However, distant control still remains poor, especially for patients with N3 disease.
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Neoplasias Nasofaríngeas/radioterapia , Radioterapia de Intensidade Modulada , Taxa de Sobrevida , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antineoplásicos/uso terapêutico , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Nasofaríngeas/tratamento farmacológico , Prognóstico , Adulto JovemRESUMO
RATIONALE: Primary spinal glioblastoma multiforme (GBM) is a rare clinical entity with an aggressive course and an invariably dismal prognosis. Its clinical characteristics, radiologic and pathologic findings, and treatment protocols have been discussed in a few cases. PATIENT CONCERNS: A 15-year-old female was admitted to the neurology department with a chief complaint of progressive numbness and weakness in her left upper extremity for 3 months and neck pain for 1 month. DIAGNOSES: Spinal magnetic resonance imaging showed an intramedullary expansile mass localized between C4 and C7. The diagnosis of GBM was determined on the basis of the histopathological findings after operation. INTERVENTIONS: Laminotomy and laminoplasty between C4 and C7 were performed, and the tumor was partially resected. The patient was administered focal adjuvant radiotherapy concomitantly with oral chemotherapy following the surgery. OUTCOMES: With severe neurologic deficits at 13 months after the diagnosis, the patient expired. LESSONS: Although therapeutic options have been improving, the prognosis of the primary spinal GBM remains poor. The treatment of primary spinal GBM entered into a central registry and multiple-center cooperation is important in establishing future therapeutic strategies.
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Vértebras Cervicais/patologia , Glioblastoma/patologia , Neoplasias da Medula Espinal/patologia , Adolescente , Terapia Combinada , Feminino , Glioblastoma/terapia , Humanos , Neoplasias da Medula Espinal/terapiaRESUMO
To examine the mechanism of ocular axial elongation in myopia, guinea pigs (age: 2-3weeks) which either underwent unilateral or bilateral lens-induced myopization (group 1) or which were primarily myopic at baseline (group 2) received unilateral intraocular injections of amphiregulin antibody (doses: 5, 10, or 15µg) three times in intervals of 9days. A third group of emmetropic guinea pigs got intraocular unilateral injections of amphiregulin (doses: 0.25, 0.50 or 1.00ng, respectively). In each group, the contralateral eyes received intraocular injections of Ringer's solution. In intra-animal inter-eye comparison and intra-eye follow-up comparison in groups 1 and 2, the study eyes as compared to the contralateral eyes showed a dose-dependent reduction in axial elongation. In group 3, study eyes and control eyes did not differ significantly in axial elongation. Immunohistochemistry revealed amphiregulin labelling at the retinal pigment epithelium in eyes with lens-induced myopization and Ringer's solution injection, but not in eyes with amphiregulin antibody injection. Intraocular injections of amphiregulin-antibody led to a reduction of lens-induced axial myopic elongation and of the physiological eye enlargement in young guinea pigs. In contrast, intraocularly injected amphiregulin in a dose of ≤1ng did not show a significant effect. Amphiregulin may be one of several essential molecular factors for axial elongation.
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Anfirregulina/farmacologia , Anticorpos/farmacologia , Miopia/prevenção & controle , Anfirregulina/administração & dosagem , Anfirregulina/imunologia , Anfirregulina/uso terapêutico , Animais , Anticorpos/administração & dosagem , Anticorpos/imunologia , Anticorpos/uso terapêutico , Cobaias , Injeções Intraoculares , Miopia/tratamento farmacológico , Retina/efeitos dos fármacos , Retina/crescimento & desenvolvimentoRESUMO
OBJECTIVE: To investigate the role of pannexin 1 channels in cisplatin-induced apoptosis in I-10 cells and the mechanisms. METHODS: MTT assay was used to assess the cytotoxicity of cisplatin (DDP) in I-10 cells. Annexin V/PI double staining and Hoechst 33258 fluorescence staining were employed to detect early- and late-stage apoptosis of the cells, respectively. Extracellular ATP level and intracellular IP3 level in the cells were detected using commercial detection kits. RESULTS: I-10 cells exposed to both CBX (a pannexin 1 channel inhibitor) and DDP showed a higher cell viability compared with the cells exposed to DDP alone (P<0.01). CBX significantly decreased cisplatin-induced early-stage apoptosis (P<0.001) and late-stage apoptosis (P<0.01), and cause obvious reductions in extracellular ATP and intracellular IP3 levels during cisplatin-induced apoptosis (P<0.05). CONCLUSION: Pannexin 1 channels participate in cisplatin-induced apoptosis in I-10 cells possibly through the ATP/IP3 pathway.
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Antineoplásicos/farmacologia , Apoptose , Cisplatino/farmacologia , Conexinas/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , HumanosRESUMO
Chemotherapeutic drug-induced apoptosis is enhanced by gap junction intercellular communication (GJIC) in a variety of tumor cells. Oxaliplatin and gefitinib are the most widely used chemotherapeutic drugs. However, the synergistic influence remains unknown in testicular cancer chemotherapy. The aim of the present study was to investigate the apoptosis induced by oxaliplatin combined with gefitinib and the potential mechanisms in I-10 testicular cancer cells. The results showed that gefitinib significantly enhanced oxaliplatin-induced apoptosis. Furthermore, the ratio of Bcl-2/Bax and the cleavage of caspase-3 and -9 were increased by gefitinib during oxaliplatin-induced apoptosis. The oxaliplatin-induced apoptosis was enhanced through the upregulation of gap junction (GJ) channels composed of connexin 43 (Cx43) by gefitinib. The mechanism of GJIC enhancement involved the suppression by gefitinib of the expression levels of Src and PKC, which phosphorylate Cx43 and reduce GJIC. PP2 (Src inhibitor) and GF109203X (PKC inhibitor) also enhanced GJIC function. Our findings demonstrated that gefitinib enhanced oxaliplatin-induced apoptosis in I-10 cells and gefitinib upregulated the GJIC by inhibiting Src and PKC-modulated Cx43 phosphorylation.
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Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Junções Comunicantes/enzimologia , Compostos Organoplatínicos/farmacologia , Quinazolinas/farmacologia , Caspase 3/metabolismo , Caspase 9/metabolismo , Linhagem Celular Tumoral , Conexina 43/metabolismo , Resistencia a Medicamentos Antineoplásicos , Ensaios de Seleção de Medicamentos Antitumorais , Sinergismo Farmacológico , Gefitinibe , Humanos , Oxaliplatina , Fosforilação , Proteína Quinase C/metabolismo , Processamento de Proteína Pós-Traducional , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína X Associada a bcl-2/metabolismo , Quinases da Família src/metabolismoRESUMO
RATIONALE: Previous studies have shown that apolipoprotein-1 (apoA-1) binding protein (AIBP) is highly associated with the regulation of apoA-1 metabolism, suggesting its role in the treatment of atherosclerosis. However, how AIBP regulates foam cell formation remains largely unexplored. OBJECTIVE: To investigate the mechanisms underlying AIBP inhibition of foam cell formation from macrophages. METHODS AND RESULTS: THP-1-derived macrophages were incubated without or with apoA-1 and AIBP, followed by assessing the formation of foam cells and the potential mechanisms. Our results showed that AIBP and apoA-1 enhanced cholesterol efflux, altered the levels of cellular free cholesterol and cholesterol ester and prevented lipid accumulation so as to reduce the formation of foam cells. Meanwhile, lack of AIBP 115-123 amino acids resulted in the loss of AIBP binding to apoA-1. Moreover, our chemiluminescent analysis showed that AIBP promoted biotin-labeled apoA-1 binding to macrophages. Besides with AIBP, more apoA-1 bound to ABCA1, a key transporter responsible for cholesterol efflux to apoA-1, as indicated by our co-immunoprecipitation assay. Our results also showed that AIBP did not regulate ABCA1 mRNA expression, but stabilized its protein from CSN2-mediated degradation. CONCLUSIONS: AIBP promotes apoA-1 binding to ABCA1 on the cell membrane of macrophages and prevents ABCA1 protein from CSN2-mediated degradation so as to prevent foam cell formation. AIBP 115-123 amino acids is at least partially responsible for its binding to apoA-1.
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Transportador 1 de Cassete de Ligação de ATP/metabolismo , Apolipoproteína A-I/metabolismo , Colesterol/metabolismo , Macrófagos/citologia , Transportador 1 de Cassete de Ligação de ATP/genética , Animais , Apolipoproteína A-I/genética , Aterosclerose/metabolismo , Transporte Biológico , Biotina/química , Complexo do Signalossomo COP9 , Linhagem Celular , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Cromatografia Líquida de Alta Pressão , Cricetinae , Células Espumosas/citologia , Humanos , Macrófagos/metabolismo , Camundongos , Ligação Proteica , RNA Interferente Pequeno/metabolismo , Proteínas Recombinantes/metabolismo , Proteínas Repressoras/metabolismo , UbiquitinaçãoRESUMO
BACKGROUND: N-stage is related to distant metastasis in nasopharyngeal carcinoma (NPC) patients. The purpose of this study was to evaluate the efficacy and toxicity of different nedaplatin-based chemotherapy regimens in advanced N2-3 stage NPC patients treated with intensity modulated radiation therapy (IMRT). PATIENTS AND METHODS: Between April 2005 and December 2009, a total of 128 patients with N2-3 advanced NPC were retrospectively analyzed. Patients were treated with IMRT concurrent with 2 cycles of chemotherapy consisting of either nedaplatin plus paclitaxel (NP group, n = 67) or nedaplatin plus fluorouracil and paclitaxel (NFP group, n = 61). Two to four cycles of adjuvant chemotherapy were then administered every 21 days following concurrent chemoradiotherapy. RESULTS: With a median follow-up of 60 months, the 5-year overall survival (OS), progression-free survival (PFS), local-regional recurrence-free survival (LRRFS), and distant metastasis-free survival (DMFS) for all patients were 81.4%, 71.5%, 87.8% and 82.0%, respectively. No significant difference in PFS (66.6% vs. 76.7%, P = 0.212) and LRRFS rates (89.0% vs. 86.3%, P = 0.664) was observed between the NP and NFP groups. The 5-year OS (75.4% vs. 88.5%, P = 0.046) and DMFS (75.1% vs. 89.0%, P = 0.042) rate were superior in the NFP group compared with the NP group. The NFP group had a higher incidence of grade 3-4 acute toxicities including bone marrow suppression (leukopenia: χ2 = 3.935, P = 0.047; anemia: χ2 = 9.760, P = 0.002; thrombocytopenia: χ2 = 8.821, P = 0.003), and both liver and renal dysfunction (χ2 = 5.206, P = 0.023) compared with the NP group. Late toxicities were moderate and no difference was observed between the two groups. CONCLUSION: IMRT concurrent with nedaplatin-based chemotherapy is an advocated regimen for patients with advanced N2-3 stage NPC. Patients with advanced N2-3 stage may be better candidates for the NFP regimen although this regimen was associated with a high acute toxicity rate.
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Quimiorradioterapia , Neoplasias Nasofaríngeas/patologia , Neoplasias Nasofaríngeas/terapia , Sobreviventes , Adulto , Antineoplásicos/uso terapêutico , Antineoplásicos/toxicidade , Carcinoma , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/diagnóstico , Estadiamento de Neoplasias , Compostos Organoplatínicos/uso terapêutico , Compostos Organoplatínicos/toxicidade , PrognósticoRESUMO
MiR-145 is downregulated and functions as a tumor suppressor in many malignancies. In this study, the biological function, molecular mechanism, and direct target genes of miR-145 in nasopharyngeal carcinoma (NPC) cells were investigated. Cell survival was detected by cell viability assay, and cell cycle was determined through flow cytometry. Invasion and migration of NPC cells were examined using cell invasion and wound healing assays, respectively. A disintegrin and metalloproteinase 17 (ADAM17) was verified as the target of miR-145 through luciferase reporter assay, qRT-PCR, and Western blot analysis. In NPC cell lines, miR-145 expression was significantly downregulated and ADAM17 protein expression was upregulated. ADAM17 was downregulated at the post-transcriptional level by miR-145 via the binding site of ADAM17-3'UTR. Transfection with miR-145 mimic suppressed cell growth and induced cell cycle arrest in the G0/G1 phase by upregulating key G0/G1 phase regulators, namely, p53 and p21. MiR-145 also inhibited cellular migration and invasion through targeting ADAM17 involving the regulation of EGFR and E-cadherin. Knockdown of ADAM17 elicited similar effects to that of miR-145 on NPC cells. This study reveals that miR-145 suppressed the invasion and migration of NPC cells by targeting ADAM17. Thus, miR-145 could be a therapeutic target for NPC.
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Proteínas ADAM/genética , Movimento Celular/genética , MicroRNAs/genética , Neoplasias Nasofaríngeas/genética , Invasividade Neoplásica/genética , Regiões 3' não Traduzidas/genética , Proteína ADAM17 , Caderinas/genética , Carcinoma , Linhagem Celular Tumoral , Regulação para Baixo/genética , Receptores ErbB/genética , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/patologia , Invasividade Neoplásica/patologia , Processamento Pós-Transcricional do RNA/genética , Fase de Repouso do Ciclo Celular/genética , Regulação para Cima/genéticaRESUMO
This retrospective study aims to examine the association of plasma Epstein-Barr virus- (EBV-) DNA levels with the tumor volume and prognosis in patients with locally advanced nasopharyngeal carcinoma (NPC). A total of 165 patients with newly diagnosed locally advanced NPC were identified from September 2011 to July 2012. EBV-DNA was detected using fluorescence quantitative polymerase chain reaction (PCR) amplification. The tumor volume was calculated by the systematic summation method of computer software. The median copy number of plasma EBV-DNA before treatment was 3790 copies/mL. The median gross tumor volume of the primary nasopharyngeal tumor (GTVnx), the lymph node lesions (GTVnd), and the total GTV before treatment were 72.46, 23.26, and 106.25 cm(3), respectively; the EBV-DNA levels were significantly correlated with the GTVnd and the total GTV (P < 0.01). The 2-year overall survival (OS) rates in patients with positive and negative pretreatment plasma EBV-DNA were 100% and 98.4% (P = 1.000), and the disease-free survival (DFS) rates were 94.4% and 80.8% (P = 0.044), respectively. These results indicate that high pretreatment plasma EBV-DNA levels in patients with locally advanced NPC are associated with the degree of lymph node metastasis, tumor burden, and poor prognosis.
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DNA Viral/sangue , Herpesvirus Humano 4/genética , Neoplasias Nasofaríngeas/sangue , Neoplasias Nasofaríngeas/virologia , Carga Tumoral , Carcinoma , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/patologia , Estadiamento de Neoplasias , PrognósticoRESUMO
Sepsis, a hyperinflammatory response that can result in multiple organ dysfunctions, is a leading cause of mortality from infection. Here, we show that orphan nuclear receptor Nur77 (also known as TR3) can enhance resistance to lipopolysaccharide (LPS)-induced sepsis in mice by inhibiting NF-κB activity and suppressing aberrant cytokine production. Nur77 directly associates with p65 to block its binding to the κB element. However, this function of Nur77 is countered by the LPS-activated p38α phosphorylation of Nur77. Dampening the interaction between Nur77 and p38α would favor Nur77 suppression of the hyperinflammatory response. A compound, n-pentyl 2-[3,5-dihydroxy-2-(1-nonanoyl) phenyl]acetate, screened from a Nur77-biased library, blocked the Nur77-p38α interaction by targeting the ligand-binding domain of Nur77 and restored the suppression of the hyperinflammatory response through Nur77 inhibition of NF-κB. This study associates the nuclear receptor with immune homeostasis and implicates a new therapeutic strategy to treat hyperinflammatory responses by targeting a p38α substrate to modulate p38α-regulated functions.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Inflamação/prevenção & controle , Lipopolissacarídeos/toxicidade , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/efeitos dos fármacos , Fenilacetatos/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/efeitos dos fármacos , Animais , Diabetes Mellitus Tipo 2/complicações , Avaliação Pré-Clínica de Medicamentos , Homeostase/efeitos dos fármacos , Inflamação/induzido quimicamente , Ligantes , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Modelos Moleculares , Conformação Molecular , Sepse/tratamento farmacológico , Sepse/genética , Fator de Transcrição RelA/antagonistas & inibidoresRESUMO
PURPOSE: To investigate the distribution of the (CCR) and its associated factors in children. METHODS: Using a random cluster sampling method, the school-based, cross-sectional Shandong Children Eye Study included children aged 4 to 18 years from the rural county of Guanxian and the city of Weihai in the province of Shandong in East China. CCR was measured by ocular biometry. RESULTS: CCR measurements were available for 5913 (92.9%) out of 6364 eligible children. Mean age was 10.0±3.3 years, and mean CCR was 7.84±0.27 mm (range: 6.98 to 9.35 mm). In multivariate linear regression analysis, longer CCR (i.e. flatter cornea) was significantly associated with the systemic parameters of male sex (P<0001; standardized regression coefficient beta: -0.08; regression coefficient B: -0.04; 95% Confidence Interval (CI): -0.05, -0.03), younger age (P<0.001; beta: -0.37; B: -0.03; 95%CI: -0.04, -0.03), taller body height (P = 0.002; beta: 0.06; B: 0.001; 95%CI: 0.000, 0.001), lower level of education of the father (P = 0.001; beta: -0.04; B: -0.01; 95%CI: -0.02, -0.01) and maternal myopia (P<0.001; beta: -0.07; B: -0.04; 95%CI: -0.06, -0.03), and with the ocular parameters of longer ocular axial length (P<0.001; beta: 0.59; B: 0.13; 95%CI: 0.12, 0.14), larger horizontal corneal diameter (P<0.001; beta: 0.19; B: 0.13; 95%CI: 0.11, 0.14), and smaller amount of cylindrical refractive error (P = 0.001; beta: -0.09; B: -0.05; 95%CI: -0.06, -0.04). CONCLUSIONS: Longer CCR (i.e., flatter corneas) (mean:7.84±0.27 mm) was correlated with male sex, younger age, taller body height, lower paternal educational level, maternal myopia, longer axial length, larger corneas (i.e., longer horizontal corneal diameter), and smaller amount of cylindrical refractive error. These findings may be of interest for elucidation of the process of emmetropization and myopization and for corneal refractive surgery.
Assuntos
Córnea/anatomia & histologia , Olho/anatomia & histologia , Erros de Refração/patologia , Adolescente , Criança , Pré-Escolar , China , Feminino , Humanos , MasculinoRESUMO
OBJECTIVES: The purpose of this study was to investigate the relationship between huntingtin-associated protein1 (HAP1) gene and radiation therapy of breast cancer cells. METHODS: HAP1 gene was transfected into breast cancer MCF-7 cells, which was confirmed by quantitative reverse transcription-polymerase chain reaction analysis (qRT-PCR) and Western blot in vitro. The changes of cell radiosensitivity were assessed by colony formation assay. Apoptosis were examined by flow cytometry. The expressions of two radiation-induced genes were evaluated by Western blot. Tumor growth was investigated in nude mice xenograft models in vivo. RESULTS: Our data showed that HAP1 gene expression was significantly increased in HAP1-transfected MCF-7 cells in comparison with the parental cells or negative control cells. The survival rate in MCF-7/HAP1 cells was significantly decreased after irradiation (0, 2, 4, 6, 8Gy), compared to cells in MCF-7 and MCF-7/Pb groups in vitro. HAP1 gene increased apoptosis in MCF-7 cells after irradiation. Additionally, the tumor volume and weight in MCF-7/HAP1+RT group were observably lower than in MCF-7/HAP1 group and MCF-7/Pb+RT group. CONCLUSION: The present study indicated that HAP1 gene expression was related to the radiosensitivity of breast cancer cells and may play an important role in the regulation of cellular radiosensitivity.
Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/radioterapia , Regulação Neoplásica da Expressão Gênica , Proteínas do Tecido Nervoso/metabolismo , Animais , Apoptose , Feminino , Citometria de Fluxo , Humanos , Células MCF-7 , Camundongos , Camundongos Nus , Transplante de Neoplasias , Proteínas do Tecido Nervoso/genética , Tolerância a Radiação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células-TroncoRESUMO
BACKGROUND: Accumulating evidence suggests that microRNA-590 (miR-590) has protective effects on cardiovascular diseases, but the mechanism is unknown. Interestingly, previous studies from our laboratory and others have shown that macrophage-derived lipoprotein lipase (LPL) might accelerate atherosclerosis by promoting lipid accumulation and inflammatory response. However, the regulation of LPL at the post-transcriptional level by microRNAs has not been fully understood. In this study, we explored whether miR-590 affects the expression of LPL and its potential subsequent effects on lipid accumulation and pro-inflammatory cytokine secretion in human THP-1 macrophages. METHODS AND RESULTS: Using bioinformatics analyses and dual-luciferase reporter assays, we found that miR-590 directly inhibited LPL protein and mRNA expression by targeting LPL 3'UTR. LPL Activity Assays showed that miR-590 reduced LPL activity in the culture media. Oil Red O staining and high-performance liquid chromatography assays showed that miR-590 had inhibitory effects on the lipid accumulation in human THP-1 macrophages. We also illustrated that miR-590 alleviated pro-inflammatory cytokine secretion in human THP-1 macrophages as measured by ELISA. With the method of small interfering RNA, we found that LPL siRNA can inhibit the miR-590 inhibitor-induced increase in lipid accumulation and secretion of pro-inflammatory cytokines in oxLDL-treated human THP-1 macrophages. CONCLUSIONS: MiR-590 attenuates lipid accumulation and pro-inflammatory cytokine secretion by targeting LPL gene in human THP-1 macrophages. Therefore, targeting miR-590 may offer a promising strategy to treat atherosclerotic cardiovascular diseases.
Assuntos
Citocinas/metabolismo , Lipídeos/análise , Lipase Lipoproteica/genética , Macrófagos/metabolismo , MicroRNAs/genética , Regiões 3' não Traduzidas/genética , Sequência de Bases , Western Blotting , Linhagem Celular Tumoral , Expressão Gênica , Células HEK293 , Humanos , Mediadores da Inflamação/metabolismo , Lipase Lipoproteica/metabolismo , Lipoproteínas LDL/farmacologia , Macrófagos/efeitos dos fármacos , Interferência de RNA , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência do Ácido NucleicoRESUMO
Chronic infections with hepatitis B virus (HBV) and/or hepatitis C virus (HCV) are the major causes of cirrhosis globally. It takes 10-20 years to progress from viral hepatitis to cirrhosis. Intermediately active hepatic inflammation caused by the infections contributes to the inflammation-necrosis-regeneration process, ultimately cirrhosis. CD8(+) T cells and NK cells cause liver damage via targeting the infected hepatocytes directly and releasing pro-inflammatory cytokine/chemokines. Hepatic stellate cells play an active role in fibrogenesis via secreting fibrosis-related factors. Under the inflammatory microenvironment, the viruses experience mutation-selection-adaptation to evade immune clearance. However, immune selection of some HBV mutations in the evolution towards cirrhosis seems different from that towards hepatocellular carcinoma. As viral replication is an important driving force of cirrhosis pathogenesis, antiviral treatment with nucleos(t)ide analogs is generally effective in halting the progression of cirrhosis, improving liver function and reducing the morbidity of decompensated cirrhosis caused by chronic HBV infection. Interferon-α plus ribavirin and/or the direct acting antivirals such as Vaniprevir are effective for compensated cirrhosis caused by chronic HCV infection. The standard of care for the treatment of HCV-related cirrhosis with interferon-α plus ribavirin should consider the genotypes of IL-28B. Understanding the mechanism of fibrogenesis and hepatocyte regeneration will facilitate the development of novel therapies for decompensated cirrhosis.